Twenty-five milliliters of YPD were inoculated with wild-type (FB2) or mutant strains (FB2ΔPma1 and FB2ΔPma2) at an initial optical density of 0.04, and cell growth was followed for 72 h by changes in optical density at 600 nm. Aliquots of 100 μL were taken at 0, 24, 48, and 72 h to determine glucose consumption using Glucose-TR kit (Spin-React, Sant Esteve de Bas, Spain). This is based on the spectrophotometric determination of hydrogen peroxide, which is produced during the reaction catalyzed by glucose oxidase.
Glucose tr kit
Manufactured by Spinreact
Sourced in Spain
The Glucose-TR kit is a laboratory equipment product manufactured by Spinreact. It is designed for the quantitative determination of glucose in biological samples.
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Lab products found in correlation
6 protocols using glucose tr kit
1
Monitoring Yeast Growth and Glucose Consumption
2
Enzymatic Hydrolysis of Wheat Straw
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Celluclast 1.5 L FG (Novozymes) was used as commercial cocktail for saccharification. An amount of Celluclast containing 0.5 U of BGLU activity was added to 66.6 mg/mL of acid slurry from wheat straw (40% content in cellulose) in 1.5 mL sodium acetate 100 mM, pH 4, at 50 °C. Samples were taken periodically and the glucose released was quantified using the Glucose TR kit from Spinreact.
Celluclast was supplemented with 0.5 U of BGLU activity from N50010 (Novozyme), TAM377 (T. amestolkiae enzymatic crude produced using glucose as carbon source) or TAM3821 (T. amestolkiae enzymatic crude produced using Avicel as carbon source).
Celluclast was supplemented with 0.5 U of BGLU activity from N50010 (Novozyme), TAM377 (T. amestolkiae enzymatic crude produced using glucose as carbon source) or TAM3821 (T. amestolkiae enzymatic crude produced using Avicel as carbon source).
3
Comprehensive Metabolic Panel Assay
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Serum triglycerides, as well as total HDL and LDL cholesterol, were assessed through conventional clinical procedures employing a multichannel Hitachi Modular analyzer (Roche Diagnostics, Indianapolis, IN, USA). Serum levels of blood urea nitrogen (BUN) and glucose were determined using the commercially available colorimetric Urea-37 kit (ref.1001325, Spinreact, Barcelona, Spain) and the Glucose-TR kit (ref.1001190, Spinreact, Barcelona, Spain), respectively. Serum levels of aspartate aminotransferase (AST/GOT) and alanine aminotransferase (ALT/GPT) were determined using the commercially available AST/GOT kit (ref. 11830, BioSystems, Barcelona, Spain) and the ALT/GPT kit (ref. 11832, BioSystems, Barcelona, Spain), respectively. Serum C-pepetide levels were determined using the commercially available ELISA kits from Millipore Co., (EZRMCP2-21K, Burlington, MA, USA). Insulin sensitivity index was computed using glucose and C-peptide concentrations in the equation: HOMA-IR C-peptide = FSG (fasting serum glucose, mM) × FSP (fasting serum C-peptide, pM)/22.5 [84 (link)].
4
Enzymatic Glucose Quantification in Cell Media
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Glucose concentration in cell medium was measured as an indirect indicator of glucose uptake by the cells. Glucose concentration was measured spectrophotometrically, using Glucose-TR kit (Spinreact, Girona, Spain), which is based on an enzymatic reaction of glucose oxidation by glucose oxidase, followed by formation of hydrogen peroxide which reacts with a chromogenic oxygen acceptor, phenol-aminophenazone in the presence of peroxidase, and the intensity of formed color is proportional to the glucose concentration in the sample. Absorbance at 492 nm was measured using an automated microplate reader (Sunrise; Tecan, Dorset, United Kingdom). Each measurement was performed in duplicate, and mean values were used for further statistical analysis.
5
Glucose Content in Liquid IC Beverages
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Free glucose content was determined in liquid IC beverages (4 mg/mL and 10 mg/mL) using the Glucose TR Kit (SpinReact, Girona, Spain) following manufacturer’s instructions. Results were expressed as mg glucose/ mL of beverage.
6
Endoxylanase Activity Quantification
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Endo-β-1,4-xylanase activity was measured by the release of reducing sugars according to the Somogyi-Nelson method (Nelson, 1944) .
One unit of endoxylanase activity was defined as the corresponding to the release of 1 μmol of reducing sugar per minute. The standard reaction mixture consisted of 2.5%
xylan, 50 mM sodium acetate (AcONa) buffer (pH 5) and the appropriate dilution of the purified enzyme or culture crude extract. Standard assays were incubated at 50 ºC and 1,200 rpm for 5 and 10 min, in order to check the linearity of the activity.
Enzymatic activity against glucose-containing substrates was measured following the release of glucose using the Glucose-TR kit (Spinreact, Vall d'en Bas, Spain) according to the manufacturer's instructions.
The BCA method was used to quantify proteins, using bovine serum albumin (BSA) as standard and Pierce reagents (Thermo scientific, Waltham, MA, USA), following the manufacturer's instructions.
One unit of endoxylanase activity was defined as the corresponding to the release of 1 μmol of reducing sugar per minute. The standard reaction mixture consisted of 2.5%
xylan, 50 mM sodium acetate (AcONa) buffer (pH 5) and the appropriate dilution of the purified enzyme or culture crude extract. Standard assays were incubated at 50 ºC and 1,200 rpm for 5 and 10 min, in order to check the linearity of the activity.
Enzymatic activity against glucose-containing substrates was measured following the release of glucose using the Glucose-TR kit (Spinreact, Vall d'en Bas, Spain) according to the manufacturer's instructions.
The BCA method was used to quantify proteins, using bovine serum albumin (BSA) as standard and Pierce reagents (Thermo scientific, Waltham, MA, USA), following the manufacturer's instructions.
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